Exposure to wood smoke is associated with various adverse health problems. Biomonitoring of smoke exposure-associated biomarkers provides accurate measurements of personally absorbed doses. As a specific metabolite of benzene, the quantitative measurement of S-phenylmercapturic acid (S-PMA) plays a vital role in evaluating human exposure to wood smoke. In this study, we developed an efficient lateral flow immunoassay (LFIA) approach for accurately and rapidly measuring S-PMA levels. Europium chelate nanoparticles (EuNPs) conjugated with purified polyclonal sheep anti-S-PMA antibodies were employed as the fluorescent detection probe. This work is based on a competitive immunoassay, where the target S-PMA competes with the immobilized antigen on the test lines for the limited antigen-binding sites on EuNP-conjugated antibodies. Due to this competition, the fluorescent intensity of the EuNPs is inversely proportional to the concentration of the target S-PMA in the sample, enabling quantitative measurement. Owing to the large Stokes shift, superior fluorescent brightness, and saturation of the EuNPs, S-PMA levels can be measured with a limit of detection of 0.32 ng/mL, a detectable range of 0.10-30 ng/mL, and a linear detection range of 0.25-30 ng/mL under optimized conditions. Stability testing revealed that the LFIA strips can be stored at room temperature for up to one year while maintaining excellent detection performance for S-PMA. These results demonstrate that the EuNP-based LFIA is a promising tool for accurate preclinical and point-of-care evaluation of wood smoke exposure. A major advantage of this approach is its ability to accurately analyze smoke biomarkers at anticipated low concentrations. The sensor system allows low-cost, rapid, and on-site data collection and quantification of wood smoke exposure.